Oligomer extended insulin-fc conjugates and their medical use

Abstract

This invention is in the field of protein conjugates. More specifically the invention relates to oligomer extended insulins with covalently attached Fc monomer polypeptides, for use in the treatment of a metabolic disorder or condition, and to methods of producing such oligomer extended insulin-Fc conjugates. The invention also relates to a novel Fc fragment, to intermediate products, and to the use of such intermediate products in processes for the synthesis of the oligomer extended insulin-Fc conjugates of the invention. Finally the invention provides pharmaceutical compositions comprising the oligomer extended insulin-Fc conjugates of the invention, and relates to the use of such compositions for the treatment or prevention of medical conditions relating to metabolic disorders or conditions.

1 . An insulin-Fc conjugate represented by Formula I: 
       
         
           
           
               
               
           
         
         wherein Ins represents an analogue of human insulin, wherein said insulin analogue is a two-chain insulin molecule comprising an A- and a B-chain, and wherein said insulin analogue comprises the amino acid substitutions A14E, A21G, B25H, B29R, and the deletion desB30;
 wherein (GQEP) 11 -GQE-(aa1) is a recombinant extension fused to the C-terminus of the insulin A-chain; and 
 wherein (aa1) is absent or proline (P). 
 
       
     
     
         2 . The insulin-Fc conjugate according to  claim 1 , wherein Ins represents an insulin analogue further comprising the amino acid substitution B16H or the amino acid substitution B16E. 
     
     
         3 . The insulin-Fc conjugate of  claim 1 , wherein Ins represents an insulin analogue selected from [A14E, A21G, B25H, B29R, desB30] human insulin, [A14E, A21G, B16H, B25H, B29R, desB30] human insulin, and [A14E, A21G, B16E, B25H, B29R, desB30] human insulin. 
     
     
         4 . The insulin-Fc conjugate of  claim 1 , wherein the oligomer extended insulin-Fc conjugate is selected from
 (A14E, A21G, A22(GQEP) 12 , A70K{circumflex over ( )}, A71P, B25H, B29R, desB30 human insulin)/(226A, 227G, 228P, 234A, 235K, 297E, 315Q, 384Q, des447 hIgG4-Fc(226-447))3,5-bis[(2-acetyl)amino]benzoyl conjugate   
       
         
           
           
               
               
           
         
         (A14E, A21G, A22(GQEP) 11 , A66G, A67Q, A68E, A69K{circumflex over ( )}, A70P, B16H, B25H, B29R, desB30 human insulin)/(226A, 227G, 228P, 234A, 235K, 297E, 315Q, 384Q, des447 hIgG4-Fc(226-447))3,5-bis[(2-acetyl)amino]benzoyl conjugate 
       
       
         
           
           
               
               
           
         
       
       and
 (A14E, A21G, A22(GQEP) 11 , A66G, A67Q, A68E, A69K{circumflex over ( )}, A70P, B16E, B25H, B29R, desB30 human insulin)/(226A, 227G, 228P, 234A, 235K, 297E, 315Q, 384Q, des447 hIgG4-Fc(226-447))3,5-bis[(2-acetyl)amino]benzoyl conjugate 
 
       
         
           
           
               
               
           
         
       
     
     
         5 . An intermediate compound Ins-(GQEP) 11 -GQE-(aa1)-KP,
 wherein Ins represents an analogue of human insulin, wherein said insulin analogue is a two-chain insulin molecule comprising an A- and a B-chain, and wherein said insulin analogue comprises the amino acid substitutions A14E, A21G, B25H, B29R, and the deletion desB30;   wherein (GQEP) 11 -GQE-(aa1) is a recombinant extension fused to the C-terminus of the insulin A-chain; and
 wherein (aa1) is absent or proline (P). 
   
     
     
         6 . The intermediate compound of  claim 5 , wherein the insulin analogue further comprises the amino acid substitution B16H or B16E. 
     
     
         7 . An intermediate compound of Formula II: 
       
         
           
           
               
               
           
         
         wherein 
         LG1 represents a leaving group reactive towards primary amino groups; and 
         LG2 represents the leaving group of a thiol reactive group. 
       
     
     
         8 . The intermediate compound of  claim 7 , wherein the intermediate compound is (2,5-dioxopyrrolidin-1-yl)-3,5-bis[(2-bromoacetyl)amino]benzoate 
       
         
           
           
               
               
           
         
       
     
     
         9 . An intermediate compound of Formula III: 
       
         
           
           
               
               
           
         
         wherein Ins represents an analogue of human insulin, wherein said insulin analogue is a two-chain insulin molecule comprising an A- and a B-chain, and wherein said insulin analogue comprises the amino acid substitutions A14E, A21G, B25H, B29R, and the deletion desB30;
 wherein (GQEP) 11 -GQE-(aa1) is a recombinant extension fused to the C-terminus of the insulin A-chain; 
 wherein (aa1) is absent or proline (P); and 
 
         wherein each LG2 represents the leaving group of a thiol reactive group. 
       
     
     
         10 . The intermediate compound of  claim 9 , wherein the insulin analogue further comprises the amino acid substitution B16H or B16E. 
     
     
         11 . An intermediate compound which is 226A, 227G, 228P, 234A, 235K, 297E, 315Q, 384Q, des447 hIgG4-Fc(226-447). 
     
     
         12 . A pharmaceutical composition comprising the insulin-Fc conjugate according to  claim 1 , and one or more pharmaceutically acceptable carriers or diluents. 
     
     
         13 . An insulin-Fc conjugate according to  claim 1  for use as a medicament. 
     
     
         14 . A method of treatment or alleviation of a disease or disorder or condition in a human, which method comprises the step of administering to such a living animal a therapeutically effective amount of an insulin-Fc conjugate according to  claim 1 , wherein said disease, disorder or condition is selected from a disease, disorder or condition relating to diabetes, impaired glucose tolerance, hyperglycemia, dyslipidemia, obesity, metabolic syndrome, hypertension, cognitive disorders, atherosclerosis, myocardial infarction, stroke, cardiovascular disorders, coronary heart disease, inflammatory bowel syndrome, dyspepsia, or gastric ulcers. 
     
     
         15 . (canceled) 
     
     
         16 . The method according to  claim 14 , wherein the disease, disorder or condition is Type 1 diabetes or Type 2 diabetes. 
     
     
         17 . The method according to  claim 14 , wherein the disease, disorder or condition is metabolic syndrome X or insulin resistance syndrome.